早孕因子(EPF)在雌性牦牛主要生殖器官中的表达  

Expression of Early Pregnancy Factors(EPF)in the Main Reproductive Organs of Female Yak(Bos grunnien)

作　　者：赵生贤 樊江峰[1] 余四九 崔燕 韩小红 马悦 何翃闳 马进彪 曹艳桃 包梅 ZHAO Sheng-Xian;FAN Jiang-Feng;YU Si-Jiu;CUI Yan;HAN Xiao-Hong;MAYue;HE Hong-Hong;MAJin-Biao;CAO Yan-Tao;Bao Mei(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep&Goat,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院,兰州730070 [2]甘肃省牛羊胚胎工程技术研究中心,兰州730070

出　　处：《农业生物技术学报》2020年第5期876-883,共8页Journal of Agricultural Biotechnology

基　　金：国家重点研发计划课题(No.2018YFD0502303);国家自然科学基金(No.31660732);甘肃农业大学科技创新基金(青年导师基金)项目(No.GAU-QDFC-2018-10);甘肃省农业科技创新课题(No.GNCX-2014-30);甘肃省高校基本科研业务费.

摘　　要：早孕因子(early pregnancy factor, EPF)是一种妊娠相关蛋白,在胚胎发育过程中作为滋养层的自分泌和旁分泌免疫耐受因子和生长因子而发挥作用。为研究EPF在雌性牦牛(Bos grunniens)主要生殖器官中的表达情况及其在繁殖活动中的作用,本研究采集牦牛卵泡期、黄体期、及妊娠早期的卵巢、输卵管、子宫以及胎盘样品,利用qRT-PCR、Western blot和免疫组化(immunohistochemistry, IHC)等技术检测其表达情况。其中IHC结果表明,在子宫腺上皮细胞、子宫内膜上皮细胞、输卵管上皮细胞、膜性黄体细胞、胎盘滋养巨细胞和隐窝上皮细胞等处均有EPF蛋白表达。qRT-PCR结果显示,卵巢中卵泡期EPF mRNA表达量极显著低于妊娠早期与黄体期;输卵管和子宫中卵泡期极显著高于黄体期与妊娠早期。妊娠早期各组织中卵巢和母体胎盘EPF mRNA表达量极显著高于其余组织。Western blot检测到EPF蛋白在卵巢和输卵管中的表达妊娠早期极显著高于卵泡期与黄体期;子宫中卵泡期表达量极显著高于黄体期与妊娠早期。在妊娠早期,卵巢与母体胎盘EPF蛋白表达量极显著高于其余器官,其中卵巢表达量最高。结果表明,EPF在雌性牦牛主要生殖器官中具有时空差异性表达特征,妊娠早期EPF可能主要来源于卵巢和母体胎盘。这一结果,为进一步研究EPF在牦牛妊娠关系的建立和维持中的作用提供基础资料。Early pregnancy factor(EPF),a kind of pregnancy related protein,plays an important role in the development of embryo as an autocrine and paracrine immune tolerance factor and growth factor of trophoblast layer.In this study,ovary,uterus,fallopian tube and placenta samples were collected respectively from female yak(Bos grunniens)during follicular phase,luteal phase and early pregnant period to investigate the expression of EPF and it role in reproduction.qRT-PCR,Western blott and immunohistochemistry(IHC)techniques were used to detect the expression of EFP mRNA and protein.The result indicated that EPF protein was most distributed in uterine glandular epithelial cells,endometrial epithelial cells,oviduct epithelial cells,membranous luteal cells,placental trophoblastic giant cells and crypt epithelial cells.qRT-PCR results showed that expression level of EPF mRNA in the ovary was significantly lower during follicular phase than that during early pregnant period and luteal phase.In oviduct and uterus the expression level of EPF mRNA was significantly higher during the follicular phase than that during the luteal phase and early pregnant period.The expression of EPF mRNA in ovarian and maternal placenta during early pregnant period was significantly higher than other reproductive organs.Western blot results showed that the expression of EPF protein in ovarian and oviduct during the early pregnant period was significantly higher than that during the luteal phase and follicular phase.In uterus of the follicular phase,the expression of EPF protein was significantly higher than that during the early pregnant period and luteal phase.It was noticeable that the expression of EPF in ovary and maternal placenta in early pregnant period was significantly higher than other reproductive organs,especially in the ovary.The results showed that EPF was spatiotemporal differently expressed in the main reproductive organs of female yak.During early pregnancy,EPF was mainly derived from ovary and maternal placenta.This study provides

关 键 词：早孕因子(EPF) 牦牛 蛋白免疫印迹 QRT-PCR 

分 类 号：S857.21[农业科学—临床兽医学]